A field experiment on “Effect of microbial consortia on growth, nodulation, yield and nutrient uptake of soybean” was carried out during Kharif 2018-19 under the Department of Soil Science & Agricultural Chemistry.The experiment was laid out under r andomized block design (RBD) with 3 replications and 9 treatments comprising beneficial microorganisms of Actinomycetes, Arthrobacter and PGPR (P. fluorescens) in possible co mbinations applied as seed treatments and additionally P.fluorescens was applied as foliar application at 20, 40 and 60 DAS of the crop growth stages.
Trang 1Original Research Article https://doi.org/10.20546/ijcmas.2019.809.306
Effect of Microbial Consortia on Growth, Nodulation, Yield and Nutrient
Uptake of Soybean in Vertisol of Central India
Bablu Yaduwanshi*, R.K.Sahu, N.G Mitra, F C Amule and Shishram Jakhar
Department of soil science and Agricultural Chemistry, Jawaharlal Nehru Krishi Vishwa
Vidhyalaya, Jabalpur, Madhya Pradesh-482004, India
*Corresponding author
A B S T R A C T
Introduction
Soybean is the most important oil seed crop in
India which contains 35-40% protein, 19%
oil, 35% carbohydrate, 5% minerals and
several other components including vitamins
(Liu, 1997) In Madhya Pradesh the area under
soybean cultivation is 54.100 lakh ha with
1,094 kg ha-1 yield and 59.170 lakh MT
production (SOPA, 2018) Soybean rhizosphere harbors vast proportions soil microorganisms, whose activities largely determine the biological condition of the soil and influence the plant growth right from seed germination to maturity (Egamberdiyeva, 2005) Among nitrogen fixing microorganisms
in soybean rhizosphere Arthrobacter is
a typical diazotroph
International Journal of Current Microbiology and Applied Sciences
ISSN: 2319-7706 Volume 8 Number 09 (2019)
Journal homepage: http://www.ijcmas.com
A field experiment on “Effect of microbial consortia on growth, nodulation, yield and
nutrient uptake of soybean” was carried out during Kharif 2018-19 under the
Department of Soil Science & Agricultural Chemistry.The experiment was laid out under r andomized block design (RBD) with 3 replications and 9 treatments comprising beneficial
microorganisms of Actinomycetes, Arthrobacter and PGPR (P fluorescens) in possible co mbinations applied as seed treatments and additionally P.fluorescens was applied as foliar
application at 20, 40 and 60 DAS of the crop growth stages The crop was supplemented with recommended dose of fertilizers 20:80:20 (N: P2O5: K2O kg ha-1) at basal application Besides these, two types of control plots were maintined as fertilized uninoculated control (FUI) and unfertilized uninoculated control (UFUI) Results revealed that the significant improvement were noticed in plant growth attributes of nodulation (57 and 43.5%), over control (9.5 and 33.4 nodule plant-1) and its biomass, (71 and 43%),over the control (0.38and 1.12 g plant-1) plant height 47, 38, 32% over the control (16.3, 35 and 45.4 cm) and its biomass, (39, 57 and 65%), over the control 1.22, 3.41and 6.07 g plant-1 leaf chlorophyll content (32 and 31.0 %) over the control (2.65 and 2.92 mg g-1leaf) all at 25,
45 and 65 DAS, nutrient contents (N, P and K seed and stover respectively) 58 and 122%,
65 and 101%, 70 and 86% over the control 98.5, 63.8, 5.2, 7.4, 24.9 and 44.4kg ha-1, yields
44 and 61% over the control 1636 and 3345 kg ha-1 respectively (seed and stover) at harvest of the crop
K e y w o r d s
soybean,
Arthrobacter,
Psuedomonas,
PGPR, nodulation,
Vertisol, FUI
Accepted:
24 August 2019
Available Online:
10 September 2019
Article Info
Trang 2The microbe is a rod shaped aerobic
Gram-positive becomes cocci at stationary growth
phase that favours the crop with nutrient
supplements including indeterminate way of N
fixation and tolerance at moisture stress
condition Some species of Arthrobacter have
been acknowledged for oxygen independent
growth strategies under limited oxygen
These species also under restriction of oxygen
use nitrate as an electron A number of
evidence has been established that PGPR
comprising Pseudomonas plays a major role in
functioning of the biosphere These bacteria
stimulate the growth of plants through direct
and indirect beneficial effects viz., enhancing
diazotrophy, nutrient solubilization,
sederophore formation for Fe availability,
excretion of growth promoting enzymes (IAA,
GA, ABA, cytokinin, etc.), vigorous growth
via ACC-deaminase activity, plants systemic
resistance (ASR and ISR(acquired and
induced systemic resistance), and
anti-oxidants (Bharadwaja et al., 2008 and Kumar
et al., 2013) acceptor at the end of their
respiratory chain, reducing it to ammonia via
nitrite
Actinomycetes are versatile group of
microorganism habitually aerobic, performing
important activities in soil like production of
growth promoting substances, phosphorus
solubilization, decomposition of organic
matter, antibiotic production for suppression
of soil borne plant pathogens etc (Franco and
Valencia, 2001)
Arthrobacter, actinomycetes and P
fluorescence individually are found beneficial
but their co-inoculation in the form of a
consortium acts as reinforced source to
augment diazotrophy, nutrient solubilization,
plant growth promoter and protectant against
soil borne pathogens especially under
moisture stress condition encountered by the
crop
Materials and Methods
The experiment was carried out during Kharif
2018 at the Research field Department of Soil Science and Agricultural Chemistry, JNKVV, Jabalpur Under the project AINP
on Soil Biodiversity & Biofertilizers (ICAR), JNKVV, Jabalpur
The recommended dose of fertilizer @ 20:80:20 kg ha-1(N: P2O5: K2O) for soybean crop was applied in the form of urea, single super phosphate (SSP) and murate of potash (MOP) SSP and MOP were supplemented as basal applications to each plot as per recommendation
One ml of gum acacia sticker solution was poured on the seed of each polythene bag followed by one ml of liquid formulations of different isolates and its consortium By holding the mouth of polythene bags seeds were enough shaken to get mixed with the sticker and inoculants formulation so that each
seed receive proper coating
Nodulation
Nodulation studies were done at 25 and 45 days of sowing by uprooting 3 plants plot-1 very carefully taking sufficient care to avoid any losses or damage of nodules After proper washing nodules of plants per plot were counted After counting, the nodules were detached from the roots and were kept in small paper bags Then the nodule fresh weight was taken Nodules were oven dried in hot air oven
at 600C for (18-20 hrs) 3-4 days (till constant weight) to record their oven dried weight
Plant height and plant dry biomass
Plant height and plant dry biomass was measured at 25, 45 and 65 DAS Three plants from each plot were taken and their heights dry weight was measured
Trang 3Chlorophyll content
Leaf chlorophyll (a, b and total) content was
estimated by acetone extraction method in
fresh plant leaves at 25 and 45 DAS
Digestion of plant samples
The plants samples were subjected to wet
digestion for estimation of various nutrients in
grain and straw Mixture of HNO3 and HClO4
was added in 2.5:1 ratio (v/v) for estimation of
major nutrients
Total nitrogen
The nitrogen content of soybean plant was
estimated on dry weight basis by
micro-kjeldahl method as per procedure suggested by
AOAC (1995)
Total phosphorus and potassium
The phosphorus contents in grain and straw of
soybean were estimated on dry weight basis
by vanado-molybdate yellow colour method
as suggested by Bhargava and Raghupathi
(1984) The potassium content in the digested
material was directly estimated Nutrients
uptake by soybean was calculated in kg ha-1 in
relation to dry with a flame photometer using
the procedure of Bhargava and Raghupathi
(1984).matter production by using the
following formula
Nutrient uptake (kg ha-1)
content (%) x yield (kg ha–1)
= -
100
Results and Discussion
Nodulation studies
The data on Nodulation of soybean is given in
Table 1 the maximum Nodulation at 25 DAS
was increased with the microbial consortium
of PGPR+Arthrobacter+ Actinomycetes
recorded the significantly higher nodules number (14.3 No plant-1) by 57% response,
followed by PGPR+ Arthro, PGPR+Actino,
Arthro+Actino, and PGPR with number of
nodules 13.7, 13.2, 12.3 and 11.7 along with response of 44.2, 42.1 29.4 and 23.2%, respectively over FUI (9.5 nodules plant-1) Similarly, the maximum number of nodules of
48 No plant-1 at 45 DAS was recorded by treatment combination of PGPR+
Arthro+Actino with 43.7% increment,
followed by PGPR+Arthro and PGPR+Actino
with nodule 41.3 and 40.8 No plant-1 by 23.65 and 22.15% response, respectively over FUI (33.4 nodules No plant-1) Jakhar et al.,
(2018) Treatment P80+LRh (80 kg P2O5 +
liquid inoculum of Rhizobium) for enhanced
the nodulation attributes, over the control at 45 DAS The treatment P80+LRh gave maximum nodules number (81.1%), weight (89.1% fresh and 78.5% dry) and N content (64.4%) over control (6.9 number/plant, 103 mg/plant, 65 mg/plant and 2.03%), respectively
The data related to nodule dry weight at 25 and 45 DAS of soybean were recorded and revealed that the consortium of PGPR+
Arthro+Actino gave the significantly higher
nodule dry wt 0.65 g with an increment 71%
over FUI (0.38 g), followed by PGPR+Arthro, PGPR+Actino, Arthro+Actino and PGPR with
nodule dry wt 0.54, 0.53, 0.51 and 0.49 g along with percent response 71, 42, 39, 34 and
29 %, respectively over FUI (0.38 g nodules plant-1) Similarly, nodule dry weight plant-1 of
soybean at 45 DAS recorded maximum with
consortium PGPR+Arthro+Actino as significantly highest (1.60 g) compared to FUI (1.12 g) by 43% response followed by
PGPR+Arthro, PGPR+Actino and Arthro+
Actino with nodule dry weight of 1.38,1.36,1.33 and 1.30 g plant-1 these treatment responded 23, 21, 19 and 16% over FUI The actinomycetes consortium (consortium of different strains of
actonomycetes) and Rhizobium consortia
along with PGPR consortia was found most
Trang 4effective among microbial inoculants, in
respect of symbiotic parameters including
nodule number, nodule dry weight
Gopalakrishnan et al., (2015).The
consortia of bradyrhizobia (Bradyrhizobium
japonicum) and phosphate solubilizing
bacteria (Pseudomonas sp.) as liquid
inoculants on soybean enhanced the nodule
number, fresh weight and dry weight
of nodules Amule et al.,(2018)
Plant height and biomass
The data on Plant height and its biomass of
soybean is given in Table 2 and 3
plant height of 23.9 cm at 25 DAS was
responded maximum with the treatment
combination of PGPR+Arthrobacter+
Actinomycetes and responded by 46.62% over
the FUI (16.3 cm) followed by PGPR+Arthro,
PGPR+Actino, Arthro +Actino with plant
height of 22.4 cm, 21.9 cm and 21.5 cm along
with 37.42, 34.29 and 31.90% response,
respectively
Similarly at 45 DAS the plant height (48.3
cm) was significantly influenced by the
treatment combination of PGPR+
Arthro+Actino with an increment 38%, over
FUI (35 cm) followed by PGPR+Arthro,
PGPR+ Actino, Arthro+Actino, with plant
height of 45 cm, 44.3 cm, 43 cm and 42.3 cm
and response increase by 28.57, 26.57, 22.85
and 20.85%, respectively
The plant height (59.8 cm) of soybean at 65
DAS exhibited of the significantly higher
plant height (59.8 cm) was found associated
with PGPR+ Arthro+Actino with an increment
31.7% over FUI (45.4 cm) This was followed
by PGPR+Arthro, PGPR+ Actino,
Arthro+Actino with plant heights of 57.5 cm,
57.1 cm, and 55.5 cm along with response
increase by 26.6, 25.7, and 22.2 %,
respectively It is well documented that soil
microorganisms exert a beneficial effect
on plant growth and development In fact, many microorganisms are being commercialized to be used in agriculture
Bashan et al., (2014)
The dry biomass 1.70 g plant-1 of soybean plant at 25 DAS was gained maximum by the
consortium PGPR+Arthro+Actino with 39%
response over the control FUI (1.22 g plant-1)
followed by PGPR+Arthro and PGPR+Actino
with plant dry biomass of 1.63 and 1.56 along with 34 and 28% response, respectively
Similarly the dry biomass at 45 DAS the
treatment combination of
PGPR+Arthro+Actino increased the plant dry
biomass by 5.14 g plant-1 with 51% response over FUI (3.41 g plant-1) followed by the
consortium PGPR+Arthro with plant biomass
of 4.52 g plant-1 by 33% increment over FUI (3.41 g plant-1)
The highest plant biomass at 65 DAS of 10 g plant-1 recorded by application of PGPR+Arthro+Actino with an increment of 65% over the control FUI (6.07 g plant-1) followed by PGPR+Arthro and PGPR+Actino with plant dry biomass of 8.86 and 8.48 g plant-1 by 43 and 40% response, respectively Jha and Saraf (2012) also reported that the root and shoot biomass were maximized with microbial consortia compared to both control and individual trials of microorganisms
Linu (2009) also confirmed the findings that the seed inoculation in cowpea by phosphate solublizers improved nodulation, root and shoot biomass, stover and seed yield and phosphorous and nitrogen uptake by crop
Co-inoculation of G fasciculatum, P fluorescens and A chrococcum enhanced shoot and root biomass of tomato (Kavatagi et al., 2014)
Chlorophyll content
The chlorophyll content in the leaf were presented in Table 4 The maximum total
Trang 5chlorophyll content (3.51 mg g-1 of leaf)was
obtained by the application of PGPR+Arthro
+Actino consortium which gives 32%
response over FUI (2.65 mg g-1 of leaf),
followed by PGPR+Arthro and PGPR+Actino
with chlorophyll content of 3.38 and 3.23 mg
g-1 of leaf by 28 and 23%
response, respectively Similarly, the data on
chlorophyll content at 45 DAS all the
treatment combinations, the consortium of
PGPR+Arthro+Actino performed significantly
better for the leaf chlorophyll content of 3.85
mg g-1 with 31% increment over the control of
FUI (2.92 mg g-1), followed by PGPR+Actino
and Arthro+Actino with total chlorophyll
content of 3.55, 3.50 and 3.45 mg g-1 of leaf
by 22, 20 and 18% response, respectively The
PGPR had profound effect on the pigment
contents on all the sampling days (25, 50, 75
and 100 DAS)
Similar finding suggested that the highest
chlorophyll „a‟, chlorophyll „b‟ and total
chlorophyll (0.805, 0.740 and 1.545 mg g-1 of
leaf) were recorded in 75 days old crop plants
grown with Rhizobium+ Pseudomonas+
Bacillus and declined in 100 DAS
(Mathivanan et al., 2017)
The PGPR (Azospirillum, Azotobacter and
Chlorophyll „a‟, Chlorophyll „b‟ and total
chlorophyll Al-Erwy et al., (2016) reported
that the A globiformis enhanced plant
biomass, uptake of iron and phosphate, protein
and chlorophyll contents Iron plays an
important role in chlorophyll biosynthesis
pathway
uptake by crop
The nitrogen, phosphorus and potassium
uptake by the soybean seed and stover were presented in Table 5 The consortium of
isolates PGPR+Arthro+Actino was recorded
maximum nitrogen uptake of 155.7 kg N ha-1
by seed and 142.0 kg N ha-1 by stover of soybean over FUI of seed (98.5 kg N ha-1) and stover (63.8 kg N ha-1)
It was interesting to note that the percent increment were computed 58 and 122%, by seed and straw, respectively next to
PGPR+Arthro, PGPR+Actino, Arthro+ Actino
and PGPR with N uptake by seed of 133.2, 130.3, 123.7 and 121.1 kg N ha-1 as well as N uptake by stover of 107, 106, 95.9 and 94.0 kg
N ha-1
Similarly, studied that the effectiveness of B
japonicum strains on soybean gave significant
positive effect on N content in shoot, N uptake
by shoot and seed with inoculation by B
japonucum or the mixed culture of strains B
japonucum Solaiman and Hossain (2006)
The consortium of PGPR+Arthro+Actino
were obtained significantly maximum P uptake of 8.6 kg P ha-1 by seed and 14.9 kg P
ha-1 by stover of soybean as compare to FUI
of seed (5.2 kg Pha-1) and stover(7.4kgPha-1) The percent responses were calculated 65 and 101%, P uptake by seed and stover,
respectively followed by PGPR+Arthro, PGPR+Actino, Arthro+Actino and PGPR with
P uptake by seed of 7.1, 7.0, 6.6 and 6.4 kg P
ha-1 as well as P uptake by stover of 11.7, 10.7, 10.5 and 10.5 kg P ha-1 due to the stimulatory effects of bacterial species such as
Pseudomonas, Bacillus and Arthrobacter were
observed on growth, yield, nitrogen and
phosphorous uptake of cotton (G hirsutum)
by Egamberdiyeva et al.,(2005)
Trang 6Table.1 Effect of microbial consortia on nodulation of soybean at different growth stages
Treatment (Nodules Plant -1 ) Dry weight ( g plant -1 )
25 DAS
sem 45
DAS
sem 25
DAS
sem 45
DAS
sem
6
0.49 0.17 1.30 0.42
5
0.53 0.05 1.36 0.38
2
0.54 0.03 1.38 0.14
F+PGPR+Arthro+Acti
no
14.3 1.15 48.0 5.13 0.65 0.08 1.60 0.17
Table.2 Effect of microbial consortia on plant height and dry biomass of
soybean at different growth stages
Treatment Plant height (cm) Plant biomass(g plant -1 )
25 DAS
DAS
sem 65 DAS sem
F+PGPR+Arthro+Acti
no
23.9 4.99 48.3 5.72 59.8 5.87
Trang 7Table.3 Effect of microbial consortia on plant dry biomass of soybean at different growth stages
Treatment Plant biomass(g plant -1 )
25DAS sem 45DA
S
sem 65DAS sem
F+PGPR+Arthro+Acti
no
1.70 1.60 5.14 2.54 10.00 1.58
Table.4 Effect of microbial consortia on chlorophyll content of soybean at
different growth stages
Treatment
Total chlorophyll content (mg g -1 leaf tissue)
25 DAS sem 45 DAS sem
Trang 8Table.5 Effect of microbial consortia on NPK uptake (kg ha-1) by seed and stover
Seed Stover Seed Stover Seed Stover
Table.6 Effect of microbial consortia on seed and stover yields of soybean
The responses of the consortium of
PGPR+Arthro+Actino were calculated 70 and
86% K uptake by seed and stover over FUI,
respectively followed by PGPR+Arthro,
PGPR+ Actino, Arthro+Actino and PGPR
with K uptake by seed of 35.2, 35.0, 33.2 and
30.9 kg K ha-1 as well as K uptake by stover of
68.9, 64.6, 63.6 and 59.8 kg K ha-1 might be
attributed due to the potential K improvement and P nutrition by application of PGPR as biofertilizers for sustainable solution to improve plant nutrient status and production (Vessey, 2003) Higher K uptake may be attributable to the mobilization of nutrients from the soil because of the secretion of organic acids mediated by soil
Trang 9microorganisms (Basak and Biswas., 2010)
Seed and stover yield of soybean
The data on grain yield and biomass
production of soybean is given in Table 6 The
grain yield of soybean differed significantly
among all the treatments The consortium of
PGPR+ Arthro+Actino gave the significantly
maximum grain yield of 2350 kg ha-1 that was
responded 44% over FUI (1636 kg ha-1)
followed by PGPR+Arthro, PGPR+ Actino,
Arthro+Actino and PGPR with grain yield of
2102, 2037, 1978 and 1947 kg ha-1 along with
the increment of 28, 25, 21 and 19%,
respectively This increment in yields of
soybean with the treatments of inoculation
fertilization might be attributed to better
nodulation, N2 fixation and crop growth as
against uninoculated control (Brahmaprakash
et al., 2004 and Gupta, 2005) Moreover, the
co-inoculation of Bradyrhizobium and
Pseudomonas strains along with P2O5
enhanced the grain yield of soybean by 38% in
pot experiments and 12% in the field
experiment, over P2O5 alone (Afzal et al.,
2010)
Similarly, the highest stover yield of soybean
(5381 kg ha-1) was recorded with the treatment
combination of PGPR+Arthro+Actino by 61%
response over the control FUI (3345 kg ha
-1
) followed by the treatment combination of P
GPR+Arthro with stover yield of 4369 kg ha
-1
by 31% which has been supported by the
findings of Amule et al., (2018) the
inoculation of microbial consortium
(actinomycetes, Rhizobium and PGPR)
supplemented with recommended dose of
fertilizer gave the significant improvement in
seed and stover yield of soybean over fertilize
d uninoculated
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